Cloned (Comment) | Organism |
---|---|
cloned into pGEX-6p-1 expression vector, expressed in Escherichia coli Bl21(DE3), glutathione S-transferase-fused CDO is purified and GTS tag is removed by cleavage with PreScission Protease | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
co-crystallization of L-cysteine and purified wild-type cysteine dioxygenase by the hanging drop, vapor-diffusion method at 16°C, the crystals grow as rods up to 0.2 x 0.2 x 0.8 mm in 1 week. Refinement of the crystal stucture leads to a final model with a crystallographic R-factor of 18.1% and a free R-factor of 21.5% at 2.7 A resolution. | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
C164A | mutation of C164 shows a about 20% abatement of enzymatic activity | Homo sapiens |
C164S | mutation of C164 shows a about 20% abatement of enzymatic activity | Homo sapiens |
C93A | C93 mutation reduces activity to 50%, zinc content of about 45%, specific activity of Cys-93 mutants is proportional to the measured iron content. | Homo sapiens |
C93S | C93 mutation reduces activity to 50%, zinc content of about 45%, specific activity of Cys-93 mutants is proportional to the measured iron content. | Homo sapiens |
R60A | similar results as R60Q, R60 mutation reduces activity to 30% | Homo sapiens |
R60Q | R60 mutation reduces activity to 30% | Homo sapiens |
Y157F | in the gel-filtration chromatography Y157F shows an additional peak with an estimated molecular weight equivalent to a cysteine dioxygenase dimer. The results for monomer and dimer are similar. Activity reduced to 5% of the wild type activity. Zinc content of about 45% | Homo sapiens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
EDTA | an EDTA:cysteine dioxygenase molar ratio of about 1000:1 abolish`s cysteine oxidase activity | Homo sapiens | |
homocysteine | cysteine dioxygenase activity is reduced by 50% only when the molar ratio of homocysteine:cysteine dioxygenase reach about 30000:1 | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
3.1 | - |
L-cysteine | under conditions where the second substrate oxygen is saturated | Homo sapiens |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Fe2+ | purified human cystein dioxygenase yields a iron incorporation of about 68% | Homo sapiens | |
Fe2+ | unsaturated distored tetragonal bipyramidal ferrous center with a vacant coordination site | Homo sapiens | |
Mg | purified human cystein dioxygenase yields trace amounts of magnesium about 0.41% | Homo sapiens | |
Mn | purified human cystein dioxygenase yields trace amounts of manganese about 0.25% | Homo sapiens | |
Zn2+ | purified human cystein dioxygenase yields a zinc incorporation of about 18.1% | Homo sapiens |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | Q16878 | - |
- |
Purification (Comment) | Organism |
---|---|
standard glutathione S-transferase fusion protein purification protocol, gel-filtration chromatography after removing the glutathione S-transferase tag | Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-cysteine + O2 | - |
Homo sapiens | 3-sulfino-L-alanine | - |
? |
Synonyms | Comment | Organism |
---|---|---|
CDO | - |
Homo sapiens |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.7 | - |
L-cysteine | determind under saturating oxygen conditions | Homo sapiens |